Rapid Identification of Carbapenemase-Producing Klebsiella pneumoniae Using Headspace Solid-Phase Microextraction Combined with Gas Chromatography-Mass Spectrometry

Background Carbapenemase-producing Klebsiella pneumoniae is an unprecedented threat to public health, and its detection remains challenging. Analysis of microbial volatile organic compounds (VOCs) may offer a rapid way to determine bacterial antibiotic susceptibility. Purpose The aim of this study was to explore the VOCs released by carbapenemase-producing carbapenem-resistant Klebsiella pneumoniae (CRKP) using headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). Methods Test bacteria were incubated in trypticase soy broth to the end of exponential growth phase, and imipenem was added in the middle time. Headspace VOCs were concentrated and analyzed using HS-SPME/GC-MS. Results The compound 3-methyl-1-butanol was found to be a biomarker among the 26 bacterial isolates (10 KPC-positive, 10 NDM-positive, 2 IMP-positive, 2 carbapenemase-negative CRKP, and 2 carbapenem-susceptible K. pneumonoiae). Conclusion This study explored a promising new strategy for the screening of carbapenemase-producing CRKP strains. Further research with larger sample sizes will potentially accelerate the application of biomarkers in routine microbiology.

[1]  N. Kasatpibal,et al.  Effectiveness and Nephrotoxicity of Loading Dose Colistin–Meropenem versus Loading Dose Colistin–Imipenem in the Treatment of Carbapenem-Resistant Acinetobacter baumannii Infection , 2022, Pharmaceutics.

[2]  Liang Chen,et al.  Multicenter Genomic Analysis of Carbapenem-Resistant Klebsiella pneumoniae from Bacteremia in China , 2022, Microbiology spectrum.

[3]  B. Buszewski,et al.  An Optimistic Vision of Future: Diagnosis of Bacterial Infections by Sensing Their Associated Volatile Organic Compounds , 2020, Critical reviews in analytical chemistry.

[4]  A. Morrin,et al.  Multi-strain volatile profiling of pathogenic and commensal cutaneous bacteria , 2020, Scientific Reports.

[5]  Suriyon Uitrakul,et al.  A Comparison of Colistin versus Colistin Plus Meropenem for the Treatment of Carbapenem-Resistant Acinetobacter baumannii in Critically Ill Patients: A Propensity Score-Matched Analysis , 2020, Antibiotics.

[6]  K. Kazmierczak,et al.  Longitudinal analysis of ESBL and carbapenemase carriage among Enterobacterales and Pseudomonas aeruginosa isolates collected in Europe as part of the International Network for Optimal Resistance Monitoring (INFORM) global surveillance programme, 2013-17. , 2020, The Journal of antimicrobial chemotherapy.

[7]  Feng Cheng,et al.  Profiles of volatile indole emitted by Escherichia coli based on CDI-MS , 2019, Scientific Reports.

[8]  B. Costello,et al.  From fast identification to resistance testing: Volatile compound profiling as a novel diagnostic tool for detection of antibiotic susceptibility , 2019, TrAC Trends in Analytical Chemistry.

[9]  N. Ratcliffe,et al.  Sniffing out resistance – Rapid identification of urinary tract infection‐causing bacteria and their antibiotic susceptibility using volatile metabolite profiles , 2019, Journal of pharmaceutical and biomedical analysis.

[10]  B. Cao,et al.  Impact of Combination Therapy vs Monotherapy on Mortality from Carbapenem-Resistant Enterobacteriaceae Bacteremia: A Retrospective Observational Study from a Chinese Network , 2018 .

[11]  J. Perry,et al.  Use of exogenous volatile organic compounds to detect Salmonella in milk. , 2018, Analytica chimica acta.

[12]  K. Bush,et al.  Past and Present Perspectives on β-Lactamases , 2018, Antimicrobial Agents and Chemotherapy.

[13]  P. Silcock,et al.  Effect of medium compositions on microbially mediated volatile organic compounds release profile , 2018, Journal of applied microbiology.

[14]  A. Goldstein,et al.  Emission Factors of Microbial Volatile Organic Compounds from Environmental Bacteria and Fungi. , 2018, Environmental science & technology.

[15]  Lisong Shen,et al.  Rapid detection of carbapenemase activity of Enterobacteriaceae isolated from positive blood cultures by MALDI-TOF MS , 2018, Annals of clinical microbiology and antimicrobials.

[16]  David K. Karig,et al.  Direct Growth of Bacteria in Headspace Vials Allows for Screening of Volatiles by Gas Chromatography Mass Spectrometry , 2018, Front. Microbiol..

[17]  P. Garbeva,et al.  Microbial Volatiles: Small Molecules with an Important Role in Intra- and Inter-Kingdom Interactions , 2017, Front. Microbiol..

[18]  Saskia Preissner,et al.  mVOC 2.0: a database of microbial volatiles , 2017, Nucleic Acids Res..

[19]  B. Buszewski,et al.  The effect of growth medium on an Escherichia coli pathway mirrored into GC/MS profiles , 2017, Journal of breath research.

[20]  B. Buszewski,et al.  Mass spectrometric techniques for the analysis of volatile organic compounds emitted from bacteria. , 2017, Bioanalysis.

[21]  Hui Shi,et al.  Characteristics of volatile organic compounds produced from five pathogenic bacteria by headspace‐solid phase micro‐extraction/gas chromatography‐mass spectrometry , 2017, Journal of basic microbiology.

[22]  J. Hill,et al.  Expanding the Klebsiella pneumoniae volatile metabolome using advanced analytical instrumentation for the detection of novel metabolites , 2017, Journal of applied microbiology.

[23]  Robert A. Weinstein,et al.  The Epidemiology of Carbapenem-Resistant Enterobacteriaceae: The Impact and Evolution of a Global Menace , 2017, The Journal of infectious diseases.

[24]  Zhongping Huang,et al.  Analysis of volatile organic compounds in pleural effusions by headspace solid-phase microextraction coupled with cryotrap gas chromatography and mass spectrometry. , 2016, Journal of separation science.

[25]  P. Nordmann,et al.  Carbapenemase-Producing Klebsiella pneumoniae, a Key Pathogen Set for Global Nosocomial Dominance , 2015, Antimicrobial Agents and Chemotherapy.

[26]  G. Peirano,et al.  The Role of Epidemic Resistance Plasmids and International High-Risk Clones in the Spread of Multidrug-Resistant Enterobacteriaceae , 2015, Clinical Microbiology Reviews.

[27]  J. Perry,et al.  Identification of volatile organic compounds produced by bacteria using HS-SPME-GC-MS. , 2014, Journal of chromatographic science.

[28]  Brendan F Gilmore,et al.  Clinical relevance of the ESKAPE pathogens , 2013, Expert review of anti-infective therapy.

[29]  Graham Bothamley,et al.  Point-of-care breath test for biomarkers of active pulmonary tuberculosis. , 2012, Tuberculosis.

[30]  Y. Liu,et al.  Outbreak of pulmonary infection caused by Klebsiella pneumoniae isolates harbouring blaIMP-4 and blaDHA-1 in a neonatal intensive care unit in China. , 2012, Journal of medical microbiology.

[31]  Aiqin Fang,et al.  iMatch: a retention index tool for analysis of gas chromatography-mass spectrometry data. , 2011, Journal of chromatography. A.

[32]  N. Woodford,et al.  Molecular mechanisms disrupting porin expression in ertapenem-resistant Klebsiella and Enterobacter spp. clinical isolates from the UK. , 2009, The Journal of antimicrobial chemotherapy.

[33]  Y. Liu,et al.  Two Clinical Strains of Klebsiella pneumoniae Carrying Plasmid-Borne blaIMP-4, blaSHV-12, and armA Isolated at a Pediatric Center in Shanghai, China , 2009, Antimicrobial Agents and Chemotherapy.

[34]  T. Hankemeier,et al.  Microbial metabolomics with gas chromatography/mass spectrometry. , 2006, Analytical chemistry.

[35]  Timothy R. Walsh,et al.  Metallo-β-Lactamases: the Quiet before the Storm? , 2005, Clinical Microbiology Reviews.

[36]  G. Jacoby,et al.  Role of β-Lactamases and Porins in Resistance to Ertapenem and Other β-Lactams in Klebsiella pneumoniae , 2004, Antimicrobial Agents and Chemotherapy.

[37]  P. Rabbitt,et al.  Long-term postoperative cognitive dysfunction in the elderly: ISPOCD1 study , 1998, The Lancet.

[38]  H. J. Kolmos,et al.  Epidemiology of Klebsiella bacteraemia: a case control study using Escherichia coli bacteraemia as control. , 1998, The Journal of hospital infection.

[39]  S. Bagley.  Habitat Association of Klebsiella Species , 1985, Infection Control.

[40]  C. D. Cox,et al.  Use of 2-aminoacetophenone production in identification of Pseudomonas aeruginosa , 1979, Journal of clinical microbiology.

[41]  O. Sicard,et al.  Monitoring of Bacterial Growth and Rapid Evaluation of Antibiotic Susceptibility by Headspace Gas Analysis , 2014 .

[42]  Scott Sutton,et al.  Measurement of Microbial Cells by Optical Density , 2011 .

[43]  P. Nordmann,et al.  Metallo-beta-lactamases: the quiet before the storm? , 2005, Clinical microbiology reviews.

[44]  G. Jacoby,et al.  Role of beta-lactamases and porins in resistance to ertapenem and other beta-lactams in Klebsiella pneumoniae. , 2004, Antimicrobial agents and chemotherapy.